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Custom Cytosine Base Editor (AAV)
Custom Cytosine Base Editor (AAV)
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$1,995
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Precision C-to-T base editing via dual-AAV split-intein delivery for in vivo applications. SpCas9-based CBE systems reconstitute in target cells from co-transduced N-terminal and C-terminal AAV vectors. Ideal for liver, muscle, and CNS applications requiring C-to-T editing with established SpCas9 PAM compatibility and editing windows. Choose BE3 for foundational validation or BE4max for enhanced efficiency and reduced indels.
Payload
U6-(gRNA)-(pro)-BE3-bGHpA: Express gRNA under U6 and BE3 under selected promoter for foundational cytosine base editing with broad validation. Uses the following parts:
- BE3 Base Editor: First cytosine base editor enabling C-to-T conversion at protospacer positions 4–8 without double-strand DNA breaks. Achieves ~30% editing efficiency with ~1.1% indels in human cells using APOBEC1 deaminase fused to Cas9 D10A nickase. Essential reference for benchmarking against optimized editors (BE4max, ABEmax) and for studying CBE mechanism and constraints.
- bGH PolyA Signal: Compact (225 bp) polyadenylation signal from bovine growth hormone with 3-fold higher expression than SV40 early polyA in key mammalian cell types. Size advantage over SV40 polyA makes it the preferred terminator for AAV vectors with limited cargo space. Standard terminator in commercial gene therapy and lentiviral vector platforms with decades of in vivo validation.
- U6 Promoter: RNA polymerase III promoter that drives constitutive expression of small RNAs. Provides strong, stable transcription with precise transcriptional start and termination sites.
U6-(gRNA)-(pro)-BE4max-bGHpA: Express gRNA under U6 and BE4max under selected promoter for enhanced cytosine editing efficiency and reduced indels. Uses the following parts:
- BE4max Base Editor: Optimized cytosine base editor with 1.8× higher efficiency than BE4. Enhanced nuclear import and codon optimization boost C-to-T conversion.
- bGH PolyA Signal: Compact (225 bp) polyadenylation signal from bovine growth hormone with 3-fold higher expression than SV40 early polyA in key mammalian cell types. Size advantage over SV40 polyA makes it the preferred terminator for AAV vectors with limited cargo space. Standard terminator in commercial gene therapy and lentiviral vector platforms with decades of in vivo validation.
- U6 Promoter: RNA polymerase III promoter that drives constitutive expression of small RNAs. Provides strong, stable transcription with precise transcriptional start and termination sites.
U6-(gRNA)-(pro)-AncBE4max-bGHpA: Express gRNA under U6 and AncBE4max under selected promoter for thermostable cytosine editing with enhanced in vivo stability. Uses the following parts:
- AncBE4max Base Editor: Advanced CBE using ancestral APOBEC reconstruction. Minimized transcriptome-wide off-targets with narrower editing window and lower toxicity.
- bGH PolyA Signal: Compact (225 bp) polyadenylation signal from bovine growth hormone with 3-fold higher expression than SV40 early polyA in key mammalian cell types. Size advantage over SV40 polyA makes it the preferred terminator for AAV vectors with limited cargo space. Standard terminator in commercial gene therapy and lentiviral vector platforms with decades of in vivo validation.
- U6 Promoter: RNA polymerase III promoter that drives constitutive expression of small RNAs. Provides strong, stable transcription with precise transcriptional start and termination sites.
U6-(gRNA)-(pro)-Target-AID-bGHpA: Express gRNA under U6 and Target-AID under selected pro for narrow-window cytosine editing with reduced bystander mutations. Uses the following parts:
- Target-AID Base Editor: Alternative cytosine base editor using sea lamprey cytidine deaminase. Narrower editing window (positions 2-4) with reduced cellular toxicity.
- bGH PolyA Signal: Compact (225 bp) polyadenylation signal from bovine growth hormone with 3-fold higher expression than SV40 early polyA in key mammalian cell types. Size advantage over SV40 polyA makes it the preferred terminator for AAV vectors with limited cargo space. Standard terminator in commercial gene therapy and lentiviral vector platforms with decades of in vivo validation.
- U6 Promoter: RNA polymerase III promoter that drives constitutive expression of small RNAs. Provides strong, stable transcription with precise transcriptional start and termination sites.
Specifications
Single-stranded AAV produced in HEK293T cells via triple transfection and purified by two rounds of CsCl ultracentrifugation. Titer determined by qPCR. Full capsid content ≥85% by mass photometry. Endotoxin ≤1 EU per 1×10¹³ VG. Purity ≥95% by SDS-PAGE. Formulated in DPBS supplemented with 150 mM NaCl and 0.001% Pluronic F-68. Store at −80°C long-term. Stable at −20°C for up to 2 months and at 4°C for 1–2 weeks after thaw.
Deliverables
- Purified AAV at 1e13 VG/mL or higher in selected volume
- Full capsid ratio 0.85 or higher
- Batch report (titer, full/empty ratio, endotoxin, SDS-PAGE)
- Payload sequence file
- Ships on dry ice
References
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